| 13C Tracer Studies of Metabolism in Mouse Tumor Xenografts
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Author:
Date:
2015-11-20
[Abstract] Mice are widely used for human tumor xenograft studies of cancer development and drug efficacy and toxicity. Stable isotope tracing coupled with metabolomic analysis is an emerging approach for assaying metabolic network activity. In mouse models there are several routes of tracer introduction, which have particular advantages and disadvantages that depend on the model and the questions addressed. This protocol describes the bolus i.v. route via repeated tail vein injections of solutions of stable isotope enriched tracers including 13C6-glucose and 13C5,15N2-glutamine. Repeated injections give higher enrichments and over longer labeling periods than a single bolus. Multiple injections of glutamine are necessary to achieve ...
[摘要] 小鼠广泛用于癌症发展和药物功效和毒性的人肿瘤异种移植研究。 稳定同位素追踪与代谢物组学分析是一种新兴的方法来测定代谢网络活动。 在小鼠模型中，有几种示踪剂引入途径，其具有取决于模型和所解决的问题的特定优点和缺点。 该协议描述了bolus i.v. 通过重复尾静脉注射稳定的同位素富集的示踪剂的溶液的路径，所述示踪剂包括13 C 13 - 葡萄糖和13 C - 13 C - ，N 15 - ，N 15 - 谷氨酰胺。 与单次推注相比，重复注射给予更高的富集和更长的标记周期。 多次注射谷氨酰胺对于在移植肿瘤中实现充分富集是必需的。
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| Assay to Evaluate Vascular Permeability Induction in Mice
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Author:
Date:
2013-11-20
[Abstract] Dengue virus infection usually courses as a benign self-limited fever, called dengue fever. However, on occasions it can progress to a life-threatening complication known as severe dengue (SD). A hallmark of SD is a sharp increase in vascular permeability. Secondary infections are considered a risk factor to develop SD, presumably through a mechanism called Antibody-Dependent Enhancement (ADE) of infection in cells with the capacity to bind antigen-antibody complexes, such as macrophages, and to trigger a subsequent aberrant cytokine response. The massive release of cytokine from macrophages has been postulated to cause changes in vascular permeability. The vascular permeability assay presented in this protocol is designed to assess whether any compound or cell-secreted product ...
[摘要] 登革热病毒感染通常称为良性自限发热，称为登革热。然而，偶尔，它可以进展为严重登革热（SD）的危及生命的并发症。 SD的标志是血管通透性的急剧增加。二次感染被认为是发展SD的风险因素，可能是通过在具有结合抗原 - 抗体复合物（例如巨噬细胞）能力的细胞中感染的感染的机制，称为抗体依赖性增强（ADE）引发随后的异常细胞因子应答。已经假定巨噬细胞中细胞因子的大量释放导致血管通透性的变化。在该方案中呈现的血管渗透性测定设计为评估来自患者的血清中存在的任何化合物或细胞分泌的产物或可溶性因子是否可以在小鼠中诱导血浆渗漏。在实验室中使用该测试以确定在存在促进性抗体的情况下由登革热病毒或登革热病毒感染的巨噬细胞在体外产生的细胞因子和可溶性因子是否能够在体内诱导血浆渗漏。在存在促进抗体的情况下，将巨噬细胞用登革病毒或登革热病毒感染48小时。此后，收集含有由巨噬细胞释放的细胞因子和可溶性因子的条件上清液，并腹膜内接种到CD-1小鼠中。在第一次接种后二十四小时，用伊文思蓝染料以第二剂量再次接种小鼠。再过24小时后，对小鼠实施安乐死，通过分光光度分析测定血液和肺中存在的伊文思蓝的量。该测定能够显示条件培养基在接种的动物中诱导血管通透性改变的能力的差异（Puerta-Guardo等人，2013）。
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| Preparation of Candida albicans Biofilms Using an in vivo Rat Central Venous Catheter Model
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Author:
Date:
2013-07-20
[Abstract] In vivo biofilms grown on medical devices are necessary to understand the interactions of the fungal biofilm and the host environment in which it is most commonly found. This protocol describes a way to grow Candida albicans biofilms on the interior lumen of central venous catheters surgically implanted into rats, which mimics quite well the clinical cases of biofilms found on human central venous catheters. These infected catheters can then be studied via a multitude of different experiments, including cell counting by plating, imaging the catheters under light or electron microscopy, or comparing the relative content of in vivo biofilms to in vitro biofilms and planktonic cultures. These biofilms also provide enough high quality RNA for ...
[摘要] 在医疗装置上生长的体内生物膜是必要的，以了解真菌生物膜和其中最常见的宿主环境的相互作用。 该协议描述了在手术植入大鼠中的中心静脉导管的内腔上生长白假丝酵母生物膜的方法，其相当好地模拟在人中心静脉导管上发现的生物膜的临床情况。 然后可以通过多种不同的实验研究这些感染的导管，包括通过电镀的细胞计数，在光学或电子显微镜下对导管成像，或者将体内生物膜的相对含量与 体外生物膜和浮游培养物。 这些生物膜还提供足够高质量的RNA进行转录分析。
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